Calcitonin gene-related peptide in Langerhans cells in psoriatic plaque lesions

Objective To study the mechanism of stress exacerbating psoriasis and the involvement effect of neuropeptides in psoriatic pathogenesis, we investigated the expression and secretion of calcitonin gene-related peptide (CGRP) in psoriatic lesions, then identified the target cells of CGRP , the characters of CGRP positive dendrite cells and the source of CGRP in psoriatic plaque lesions. Methods Specific radioimmunoassay (RIA) and immunohistochemistry staining methods were used to determine CGRP secretive content and the target cells of CGRP in psoriatic plaque lesion tissue of vulgaris psoriasis. Double immunofluorenscence staining was done on psoriatic plaque lesion sections by first using rabbit anti-human CGRP antibody and mouse anti-human CD1a antibody, second using PE-conjugated anti-mouse immunoglobulin and FITC-conjugated anti-rabbit immunoglobulin. Confocal laser microscope showed the psoriatic lesion sections. Then both digoxigenin labelled anti-sense and sense RNA probe of CGRP were synthesized to make sure the source of CGRP on the dendrite cells. The psoriatic lesion sections were studied by in situ hybridization. Results The content of CGRP in vulgaris psoriatic plaque lesions was higher than that of normal controls (P<0.01). CGRP was also found on the dermal microvascular endothelial cells and the epidermal dendrite cells in psoriatic plague lesions. Further study showed that CGRP existed on the surface of epidermal CD1a + Langerhans cell in psoriatic plaque lesion. The CGRP mRNA expressed around the nucleus of the Langerhans cells in psoriatic lesion. Conclusions The pathogenesis of psoriatic plaque lesions was closely related to the overexpression of neuropeptide CGRP. The CGRP contacted with the dermal microvascular endothelial cells and epidermal dendrite cells in psoriatic plaque lesion. The CGRP positive epidermal dendrite cell was CD1a + Langerhans cell. The Langerhans cell itself expressed CGRP mRNA.     

Calcitonin gene-related peptide in Langerhans cells in psoriatic plaque lesions

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降钙素基因相关肽与银屑病Langerhans细胞的相关性

目的:探讨神经递质降钙素基因相关肽(calcitonin gene-related peptide, CGRP) 与银屑病皮损Langerhans细胞的相关性,研究银屑病的发病机制.方法:抗生物素蛋白-生物素-过氧化物酶复合物法确定CGRP的靶细胞,用双相间接免疫荧光染色法,把CD1a单克隆抗体和CGRP抗体作用于寻常型银屑病斑块型皮损同一切片,用PE-αM IgG和FITC-αR IgG 抗体显示,通过激光扫描共焦显微镜观察,并利用RNA探针原位杂交技术,研究表皮树枝状细胞CGRP的来源.结果:免疫组化染色银屑病皮损显示CGRP阳性表皮树枝状细胞,经双相免疫荧光染色及激光扫描断层光切,可见CGRP存在于银屑病皮损表皮CD1a+ Langerhans细胞膜表面,Langerhans细胞内有CGRPmRNA的表达.结论:寻常型银屑病斑块型皮损表皮Langerhans细胞与神经肽CGRP密切接触,同时Langerhans细胞内有CGRPmRNA的表达.     

白细胞介素10在银屑病中的表达特征及影响

目的：分析银屑病患者及银屑病小鼠模型皮损处白细胞介素10(IL-10)特点及其与银屑病严重程度的相关性,探讨IL-10的表达对银屑病的影响。方法：收集2018年10月至2019年12月温州医科大学附属第二医院育英儿童医院收治的40例银屑病患者病损皮肤,同时通过咪喹莫特(IMQ)分别诱导C57BL/6(WT)小鼠和IL-10敲除(IL-10^-/-)小鼠致银屑病模型。采用PASI评分观察皮损动态变化;HE染色观察皮损组织形态学变化、表皮厚度及炎症细胞浸润程度;免疫组化检测皮损组织中IL-10和CD19蛋白表达;免疫荧光染色分析皮损组织中CD4定位改变情况。结果：IMQ造模可以取得与临床银屑病患者相类似的皮肤表现。IL-10^-/-造模组小鼠背部皮损处红斑评分、鳞屑评分、皮肤浸润厚度评分及PASI总评分和耳朵厚度较WT造模组和IL-10^-/-对照组均明显升高(P<0.05)。免疫组化结果显示WT造模组小鼠皮损组织中IL-10的表达阳性率低于WT对照组,差异有统计学意义(P<0.05)。IL-10^-/-     

The Expression of Interleukin-22 and S100A7, A8, A9 mRNA in Patients with Psoriasis Vulgaris

In order to study the expression of interleukin-22 （IL-22） and S 100A7, A8, A9 mRNA in the skin lesions of patients with psoriasis vulgaris and their relationship, the biopsies were taken from skin lesions in 35 patients with psoriasis vulgaris and the skin of 16 normal controls, and the expression levels of 1L-22 and S 100A7, A8 and A9 mRNA were detected by semi-quantitative RT-PCR. The results showed that （1） IL-22 and S 100A8, A9 mRNA were positively expressed in the psoriatic skin lesions but negatively expressed in the normal controls; The expression level of S 100A7 was （1.133±0.040） in the psoriatic skin lesions, significantly higher than that in the normal controls （0.744±0.037, P〈0.01）. （2） There were significantly positive correlations between the expression of IL-22/S100A7 mRNA, IL-22/S100A8 mRNA, IL-22/S100A9 mRNA in the psoriasis vulgaris （r1=-0.543, r2=0.774, r3=0.621, P〈0.01）. It was concluded that IL-22 and S 100A7, A8, A9 might play important roles in the occurrence and progression of psoriasis.     

银屑病患者皮损间充质干细胞对HaCaT细胞增殖的影响

目的探讨银屑病患者皮损间充质干细胞（mesenchymal stem cells,MSCs）对HaCaT细胞增殖的影响,揭示银屑病可能的免疫发病机制。方法寻常性银屑病患者8例,进行期4例,静止期4例,银屑病皮损面积和严重程度（PASI）评分范围为1．6—18．0,平均11．64。健康对照组8例,取自我院泌尿外科和整形外科手术切除的正常皮肤。分离、培养患者皮损与健康人皮肤MSCs,流式细胞术进行细胞鉴定;将第5代皮肤MSCs与HaCaT细胞共培养,并设自然增殖组,采用实时细胞分析系统进行HaCaT细胞增殖检测,培养74h后用细胞计数法检测HaCaT细胞的数量。结果倒置显微镜下,银屑病组与健康对照组皮肤MSCs的细胞形态相似,细胞表面抗原均高表达CD29、CD44、CD73、CDg0、CDl05,而CD34、CD45及HLA—DR表达阴性。皮肤MSCs可抑制HaCaT细胞的增殖（P〈0．05）,与健康对照相比,银屑病患者皮损MSCs抑制HaCaT细胞增殖的能力显著减弱[患者组细胞数为（2．35±0．254）×105,对照组细胞数为（2．04±0．122）×105,P〈0．05]。结论银屑病患者皮损MSCs抑制角质形成细胞增殖的能力减弱,这可能是银屑病的发病机制之一。     

寻常型银屑病患者皮损中IL-22与IL-22R1、STAT3、c-myc的mRNA表达及关系研究

目的探究白细胞介素-22(IL-22)及IL-22R1、信号转导和转录激活因子-3(STAT3)、原癌基因(c-myc)在银屑病表皮异常增殖中的作用。方法采用逆转录聚合酶链反应(RT-PCR)技术对23例寻常型银屑病患者皮损和11例正常人皮肤中IL-22、IL-22R1、STAT3、c-myc的mRNA水平进行检测。结果 IL-22、IL-22R1、STAT3、c-myc的mRNA在寻常型银屑病皮损中和正常人皮肤中均呈阳性表达;但在寻常型银屑病皮损中的表达水平较正常人组明显升高,差异有统计学意义(P0.01)。IL-22的mRNA水平与IL-22R1、STAT3的mRNA水平呈正相关(P0.05);STAT3的mRNA水平与c-myc的mRNA水平呈正相关(P0.05)。结论 IL-22可能诱导STAT3活化导致银屑病角质形成细胞(KC)过度增殖,这为银屑病治疗提供了新的靶点。     

T-lymphocyte chemotaxis to IL-8 in patients with psoriasis in vitro

OBJECTIVE: The critical role of infiltrating T cells in the pathogenesis of psoriasis is now well established. In order to determine whether circulating T cells from patients with psoriasis were also involved in the disease process, the authors investigated the biological behavior as studied by chemotactic activity of T cells in patients with psoriasis. METHODS: A 48 microchemotaxis chamber was employed to determine T-cell chemotaxis activity. In addition, the expression of T cell activation markers such as HLA-DR and interleukin 2 receptor were analysed with fluorescence activated cell sorting technique and serum IL-8 level was measured with ELISA methods. Forty-five patients with psoriasis (23 patients with severe psoriasis and 22 with mild psoriasis) and 21 patients with atopic dermatitis were investigated. For comparison, T-lymphocytes from 20 healthy controls were tested equally. RESULTS: T-cell chemotactic responses were significantly decreased in patients with severe psoriasis and atopic dermatitis as compared to healthy controls. Increased expression of activation markers such as HLA-DR and interleukin 2 receptor were demonstrated in circulating T cells from psoriatic patients and atopic dermatitis patients in comparison to healthy controls. Serum IL-8 level was significantly increased in patients with psoriasis and atopic dermatitis. CONCLUSIONS: Circulating T cells in patients with severe psoriasis show abnormal in vitro chemotactic response to IL-8. Furthermore, the in vivo activation state of T lymphocytes in these patients and increased level of serum IL-8 seemed to be associated to their decreased in vitro T-cell chemotactic responses.     

Caspase3、Bcl-2和PCNA在银屑病皮损组织中的表达

目的　探讨银屑病发生发展中细胞增殖和凋亡相关指标表达的趋势。方法　应用免疫组化技术检测 30例寻常型银屑病皮损、邻近皮肤及 10例正常皮肤中PCNA、Bcl - 2、Caspase3的表达。结果　在正常表皮中Bcl - 2和PCNA在基底层阳性表达 ,但在寻常型银屑病皮损中Bcl- 2表达明显低于正常皮肤组 ,皮损处与邻近皮损处表达、邻近皮损处与正常皮肤表达比较有统计学差异 (均P <0 0 5 ) ;PCNA在银屑病皮损处为角质形成细胞广泛阳性表达 ,皮损处与邻近皮肤的表达、邻近皮肤与正常皮肤相比有统计学差异 (P <0 0 1) ;Caspase3在寻常型银屑病皮损处与邻近皮肤及正常皮肤的表达无统计学差异 (P >0 0 5 )。结论　银屑病时 ,PCNA过度表达 ,Bcl - 2低表达 ,表明细胞过度增殖 ,凋亡基因Caspase3表达无统计学差异     

Study on the Expression of IL-18, AcPL and IL-18BP mRNA in the Psoriatic Lesions

Interleukine- 1 8(IL- 1 8) isa newly identified cy-tokine that plays an important role in the T- helper(Th1 ) response,primarily via its ability to induceIFN- γ production in T cells and NK cells.IL- 1 8isrelated to the IL- 1 family in terms ofstructureand interms of function,and induction of IFN-γ by IL- 1 8is similar to that by IL- 1 2 in thatthey are a sole cy-tokine,and there is synergism between IL- 1 8andIL - 1 2 for IFN-γ production[1— 4 ] .In addition,mono-cytes and macro…     

银屑病角质形成细胞抗菌蛋白的表达水平比特应性皮炎高

最近，研究显示特应性皮炎患者皮损的抗菌肽表达水平比银屑病患者低。本文采用微阵列检测方法，分析慢性斑块性银屑病和慢性特应性皮炎患者纯化的皮损处表皮细胞的mRNA表达谱，研究是否为宿主防御蛋白的普遍现象，并探讨其特异性。选择一组基因用定量PCR和免疫组化证实微阵列的数据。研究发现，相对于特应性皮炎皮损，银屑病皮损角质形成细胞有许多抗菌蛋白过度表达。有意思的是，正常分化和激活／过度增殖表皮表型的标志物表达水平一致。就细胞增殖而言，银屑病和特应性皮炎患者的慢性皮损惊人相似。研究推断银屑病表皮宿主防御蛋白的表达水平比特应性皮炎表皮高，这种现象对这些蛋白是特异性的。这是由导致表皮抗菌反应的基因多态性所致还是由银屑病与特应性皮炎细胞浸润的差别引起，仍需进一步研究。     

A study on the expression of interleukin(IL)-10 and IL-12 P35, P40 mRNA in the psoriatic lesions

Psoriasisisa common chronic,relapsing and in-flammatory disease and its pathogenesis is still un-known.It is considered thatit is one kind of geneticand immunological diseases controlled by multiplegenes.Even though theexactpathogenesisisstillun-known,many studies showed that T cells mediatedimmune response played a very important role in thepathogenesis of psoriasis.In this study,the expres-sion of IL- 1 0 ,IL- 1 2 P35 and P40 m RNA in the pso-riasis lesions were detected by using RT- P…     

银屑病患者外周血单个核细胞白介素20及其受体mRNA水平检测

目的：观察银屑病患者外周血单个核细胞（PBMC）中白细胞介素20（IL-20）及其受体（IL-20R）mRNA水平,探讨其与银屑病发生发展的关系。方法：提取外周血总RNA,应用半定量逆转录-聚合酶链反应,以β-actin为内参照,检测IL-20和IL-20R mRNA。结果：银屑病患者PBMC分泌的IL-20水平低于对照组,而IL-20R水平高于对照组。结论：推测IL-20参与银屑病的发病机制,可能是PBMC分泌的IL-20一部分与PBMC自身的受体结合,调节PB-MC自身的免疫功能;另一部分定向转运到皮肤,与其中的受体结合,从而激活Stat信号通路,进一步激活一些与机体炎症反应密切相关的基因,参与银屑病患者机体免疫调节。     

解毒药与凉血药配伍对咪喹莫特诱导的银屑病样小鼠模型的干预作用

目的 观察凉血解毒汤与凉血汤对咪喹莫特诱导的银屑病样小鼠模型皮损的影响,并探讨配伍解毒药在银屑病中的作用机制。方法 BALB/c小鼠55只,背部剃毛,采用数字表法随机分为空白对照组、模型组、凉血汤组、凉血解毒汤组和甲氨蝶呤组,采用5%（质量分数）咪喹莫特乳膏背部涂抹诱导皮肤银屑病样模型。观察皮损面积和疾病严重程度（psoriasis area and severity index,PASI）并评分,光镜下观察皮损组织形态学变化及表皮层厚度;采用皮肤水分油分测试笔检测小鼠背部皮肤水分和油分含量;采用免疫组织化学染色法观察小鼠皮损中增生细胞核抗原（proliferating cell nuclear antigen,PCNA）和T淋巴细胞表面标志CD3表达;采用RT-PCR技术检测皮损中白细胞介素-17（interleukin-17,IL-17）、IL-23及IL-1β mRNA表达。结果 与模型组相比,凉血解毒汤和凉血汤组小鼠皮损PASI评分及表皮厚度均明显降低、PCNA阳性细胞百分比下降、CD3阳性T细胞表达个数减少、IL-1β mRNA相对表达量相对下降（P<0.05）;此外凉血解毒汤还可使小鼠皮损中水分（16.980±2.739）、油分（7.750±1.209）含量有效升高（P<0.05）,IL-17、IL-23 mRNA相对表达量明显下降（P<0.05）,对皮损PASI评分及表皮厚度、PCNA阳性细胞百分比（5.66±0.057）和CD3阳性T细胞表达个数（10.670±0.193）的影响也更为显著（P<0.01）。结论 凉血解毒汤与凉血汤均可抑制IL-1β mRNA的表达,减少炎性浸润,改善咪喹莫特诱导的银屑病样炎性反应皮损;而配伍解毒药的凉血解毒汤,则可进一步通过抑制IL-23、IL-17的表达,提升其在改善皮肤屏障功能及皮损组织形态变化、减缓表皮角质形成细胞增生、减少角化不全细胞及降低炎性反应浸润程度等方面的干预作用,从而缓解咪喹莫特诱导的银屑病样皮损改变,提示解毒中药对炎性反应及相关因子的抑制作用可能是其治疗银屑病的潜在机制之一。     

寻常型银屑病患者皮损中HBD-2、IL-1β和NF-κBp65表达的研究

目的：探讨HBD-2、IL-1β和NF-κBp65在寻常型银屑病皮损中的表达及其在银屑病中的作用机制。方法：采用SP免疫组化法检测45例银屑病患者皮损区、非皮损区及15例正常人皮肤中HBD-2、IL-1β和NF-κBp65的表达,对其在皮损中的表达进行相关性分析,并将结果与PASI评分进行相关性分析。结果：银屑病患者皮损区HBD-2、IL-1β、NF-κBp65的表达比非皮损区、正常人皮肤明显上调,非皮损区HBD-2、IL-1β的表达也高于正常人皮肤,差异均有统计学意义（均P〈0．05）。银屑病患者皮损中HBD-2、IL-1β和NF-κBp65的表达水平与PASI评分之间均存在正相关（r=0．867、0．865、0．821,均P〈0．05）。银屑病患者皮损中HBD-2与IL-1β、HBD-2与NF-κBp65、IL-1β与NF-κBp65表达水平之间均存在正相关（r=0．869、0．740、0．814,均P〈0．05）。结论：HBD-2、IL-1β、NF-κBp65在寻常型银屑病皮损中的表达均升高．可能共同参与银屑病的发病过程。     

IL-15在寻常型银屑病患者皮损中的表达

目的从mRNA及蛋白质水平研究白介素15（IL-15）在寻常型银屑病患者皮损及非皮损皮肤中的表达,探讨其在银屑病发病中的作用。方法采用RT-PCR和免疫组化法对28例寻常型银屑病患者皮损及非皮损皮肤中IL-15的表达情况进行了检测,同时以10例正常人皮肤作为对照。结果2种IL-15前体蛋白的mRNA在所有皮肤标本中都有表达,银屑病皮损中IL-15 mRNA的表达明显高于非皮损处皮肤（P〈0．001）和正常人皮肤（P〈0．001）,非皮损皮肤的表达与正常人皮肤相比差异无显著性（P〉0．05）。免疫组化结果显示：IL-15低表达于非皮损和正常皮肤的基底细胞层,而在银屑病皮损处高表达,且表达范围可达表皮全层及真皮乳头层。皮损处IL-15的表达明显高于非皮损（P〈0．001）和正常皮肤（P〈0．001）,非皮损与正常皮肤之间的表达无显著性差异（P〉0．05）。结论寻常型银屑病的皮损中高表达IL-15,在疾病的发生中可能起一定的作用。     

金施尔康对银屑病病人外周血T淋巴细胞亚群及sIL—2R的影响

①目的　探讨金施尔康对银屑病病人T淋巴细胞亚群及可溶性白细胞介素 2受体 (sIL 2R)水平的影响。②方法　分别用碱性磷酸酶抗碱性磷酸酶法 (APAAP)和双抗体夹心ELLSA法 ,检测 43例寻常型银屑病病人金施尔康治疗前后外周血CD+ 4,CD+ 8,sIL 2R水平 ,同时选 35例正常人作对照组。③结果　银屑病病人血清CD+ 8,sIL 2R明显增高 ,而CD+ 4,CD+ 4/CD+ 8明显降低 ,与对照组比较差异有显著性 (t =2 .733.2 6 ,P <0 .0 1) ;应用金施尔康治疗后 ,银屑病病人CD+ 4,CD+ 4/CD+ 8明显增高 ,而CD8,sIL 2R及PASI积分明显降低 ,与治疗前比较差异有显著性 (t=2 .8910 .15 ,P <0 .0 1)。④结论　应用金施尔康治疗寻常型银屑病可明显影响病人的T淋巴细胞功能及血清sIL 2R水平。     

HIF-1α与iNOS在银屑病皮损中的表达 及其与血管生成的关系

目的：探讨缺氧诱导因子-1α(HIF-1α)和诱导型一氧化氮合酶（iNOS）在银屑病皮损组织中的表达并分析两者的相关性,同时研究两者在银屑病血管形成中所起的作用。方法：采用免疫组织化学SABC法和Western 印迹检测32例银屑病皮损组织和20例正常人皮肤组织中HIF-1α与iNOS蛋白的表达;同时用CD34标记血管内皮细胞,并计算微血管密度(MVD) 。结果：20例正常对照组表皮组织HIF-1α和iNOS表达弱或几乎无表达;32例银屑病患者皮损组织中HIF-1α和iNOS均为强表达,且与正常对照组相比,差异有统计学意义(P＜0.05)。银屑病患者皮损组织中HIF-1α和iNOS的表达均与MVD值呈明显正相关(分别为r=0.743, P＜0.01;r=0.639, P＜0.01);银屑病皮损组织中iNOS的表达与HIF-1α的表达呈明显正相关(r=0.717, P＜0.01)。结论：银屑病皮损组织存在HIF-1α和iNOS蛋白的过表达,可能通过促进银屑病新生血管的生成,对银屑病的发生发展起重要作用。